Selection and validation of suitable reference gene for qPCR gene expression analysis in lamb testis cells under Sheep pox virus infection.

Virus infection alters host gene expression, therefore ideal and stable reference housekeeping genes are required to normalise the expression of other expressed host genes in quantitative real-time PCR (qRT-PCR). The suitable reference gene may vary in response to different viral infections in different hosts or cells. In the present study, we cultured primary lamb testis cells (LTC) and assessed the expression stability of seven widely used housekeeping genes (B2M, HMBS, HPRT1, HSP-90, POLR2A, 18s_RNA, GAPDH) as reference genes in Sheeppox virus (SPPV) infected and control (uninfected-0h) LTC at 0.5h, 4.0h, 8.0h, and 12.0h post-infection) using NormFinder, Bestkeeper, geNorm, and the comparative ΔCT method in RefFinder based on their expression levels. Analysis revealed that HSP90, 18s_RNA, HPRT, POLR2A, and B2M were the most stable genes from the panel in the individual analysis group in 0h, 0.5h, 4.0h, 8.0h, and 12.0h, respectively. Furthermore, B2M was shown to be the most stable reference gene in the combined control with the respective and overall infected groups, except the control group of 4.0hpi of SPPV infection. In this study, we selected the most suitable reference genes in LTC for particular time points of SPPV infection. The identified most suitable housekeeping gene can be used during normalization of expression of other targeted genes at aspecific time point of SPPV infection.

Genome-wide expression analysis reveal host genes involved in immediate-early infections of different sheeppox virus strains.

This study explored the transcriptome of lamb testis cells infected with sheeppox virus (SPPV) wild strain (WS) and vaccine strain (VS) at an immediate-early time. Most of the differentially expressed genes (DEGs) and differentially expressed highly connected (DEHC) gene network were found to be involved in SPPV-VS infection compared to SPPV-WS. Further, the signaling pathways were mostly involved in SPPV-VS infection than SPPV-WS. SPPV modulates the expression of several important host proteins such as CD40, FAS, ITGß1, ITGα1, Pak1, Pak2, CD14, ILK leading to viral attachment and entry; immune-related DEGs such as MAPK, JNK, ERK, NFKB, IKB, PI3K, STAT which provide optimal cellular condition for early viral protein expression; and FOXO3, ATF, CDKNA1, TCF, SRF, BDNF which help in inducing apoptosis and MPTP, BAD and Tp53 inhibits apoptosis or cell death at the immediate-early time. The results captured the specific genes and enabled to understand distinct pathogenic mechanisms employed by VS and WS of SPPV.

Endoscopic single stage trans-oral decompression and anterior C1 lateral mass and C2 pedicle stabilization for atlanto-axial dislocation.

Although most of the cases of atlanto-axial dislocation (AAD) and basilar invasion can be managed by posterior approaches in the recent times, anterior decompression with stabilization is required in selected patients who persist with irreducible AAD even after manipulation of the C1-C2 facet joint under general anesthesia. A single stage endoscopic trans-oral decompression and stabilization can be used in such patients. It has not been described so far to the best of authors' knowledge. This is indicated in irreducible AAD with the mandibular angle lying below the C2-C3 disc space. It is not a proper choice when the mandibular angle is above the C2-C3 disc space, there is involvement of the facet joint by trauma or any other pathologies, and if a posterior compression at the cervicomedullary junction persists. All patients should undergo pre-operative radiographs, computed tomography (CT) scan and magnetic resonance imaging with angiogram of the cranio-vertebral region. Utilizing this technique, an intra-operative satisfactory reduction of the dislocation with C1-C2 stabilization could be achieved in 3 patients, and 7 required an additional odontoid excision. Post- operative plain radiographs should be performed to assess for C1- C2 alignment and fusion at 3 and 12 months after surgery. All 10 patients of our series had an irreducible AAD and two had an additional basilar invasion. All patients improved from the pre-operative Ranawat grade 3A (n = 8) and 3B (n = 2) to post-operative grade 1 (n = 9) and 2 (n = 1) at a 3-12- month follow-up assessment. The average duration of the procedure and blood loss was 145 minutes and 75 ml, respectively. Endoscopic trans-oral single stage decompression and stabilization seems to be an effective and safe alternative in selected patients with AAD and basilar invasion.

Endoscopic surgery of spontaneous basal ganglionic hemorrhage.

INTRODUCTION: Although there are controversies about the optimal management of spontaneous intracerebral hemorrhage (ICH), benefits of endoscopic procedures in ICH have been reported. This study is aimed to evaluate the result of 270 patients undergoing endoscopic treatment of ICH. METHODS: This was a retrospective study from July 2008 to June 2017. All procedures were done with the endoscopic technique using a tubular retractor. Patients with the hematoma volume between 30 to 80 ml, with the Glasgow Coma Scale (GCS) between 5 to 14, and evidence of severe mass effect, were included in the study. RESULTS: The average stay in the intensive care unit was 6 days (range 1-17 days). The median pre-operative midline shift of 8.3 mm was reduced to 2.7 mm after surgery. The average hematoma removal ratio, the duration of surgery, and the blood loss was 90%, 90 min, and 60 ml, respectively. There was improvement in the average pre-operative GCS from 9.4 to 11.3 at seventh post-operative day. The post-operative mortality rate was 10.7%. A good outcome was observed in 71% patients at 6 months after surgery. Larger the volume of hematoma, more the operative time, more the pre-operative midline shift, and poorer the GCS, significantly higher was the association with mortality. The patients with a better pre-operative GCS were associated with a better Glasgow Outcome Score. The follow-up period ranged from 7 to 115 months. CONCLUSION: Endoscopic surgery with the help of a tubular retractor was effective and safe. It allowed for a good visualization of the hematoma and the surrounding brain, and helped in proper hemostasis. The hematoma may also be removed with the help of the microscope and the tubular retractor, in case any difficulty during the endoscopic technique is encountered.

Delayed Paraplegia in an Adult Patient With Spinal Cord Injury without Radiographic Abnormality of Dorsal Spine: A Lesson Learned.

Noncompressive myelopathy of lower dorsal spine secondary to trauma is a rare event. We report a case of delayed paraplegia in a patient with a history of road traffic accident. The X-ray of dorsolumbar spine did not show any abnormality. Magnetic resonance imaging of dorsolumbar spine was performed which showed the presence of central T2-weighted hyperintensities from D10-D11 to D12-L1 level. No associated bony injury was documented, and the integrity of the spinal canal was maintained. The patient was managed conservatively with bed rest, and steroids were given. However, the patient did not show any signs of improvement after 1 month of follow-up.

Endoscopic Anterior Approach for Cervical Disc Disease (Disc Preserving Surgery).

OBJECTIVE: To report our experience of endoscopic disc removal by anterior approach for management of cervical myelopathy in 210 patients. METHODS: A retrospective study of 187 cases of single- and 23 cases of double-level disc disease was performed. Cases of myelopathy with or without unilateral or bilateral radiculopathy and unilateral radiculopathy with either soft or hard disc prolapse were included. Patients with ≥3 disc levels, unstable spine, infections, trauma, significant posterior compression, congenital canal stenosis, disc extending more than half the vertebral body height, and prior surgery at the same level were excluded. RESULTS: C5-6 (n = 119 patients), C6-7 (n = 58 patients), C4-5 (n = 49 patients), C3-4 (n = 6 patients), and C2-3 (n = 1 patient) levels were represented. Visual analog scale and Nurick grading system were used to assess severity of neck and arm pain and functional outcomes, respectively. Preoperative mean visual analog scale scores for arm and neck pain were 6.7 and 3.2, respectively, which improved to 1.7 and 1.1 at 3 months after surgery. The average preoperative Nurick grade improved from 2.64 to 0.81 at 6 months postoperatively. Follow-up was 6-54 months. CONCLUSIONS: Endoscopic anterior discectomy (disc preserving surgery) is an effective and safe alternative in cervical disc disease. Although there was reduction in disc height, clinical outcome was good at an average 29 months of follow-up. Long-term follow-up is required to assess any progressive disc degeneration and clinical results.

Role of Diffusion Tensor Imaging in Brain Tumor Surgery.

OBJECT: The objective is to evaluate the role of diffusion tensor imaging (DTI) in intra-axial brain tumor cases (gliomas and metastasis). To preoperatively assess the integrity and location of white matter (WM) tracts and plan the surgical corridor to cause least damage to the WM tracts with minimum postoperative new neurological deficits. MATERIALS AND METHODS: A total of 34 patients were included in this study. Pre-operative contrast-enhanced magnetic resonance imaging and DTI scans of the patients were taken into consideration. Pre- and post-operative neurological examinations were performed and the outcome was assessed. RESULTS: Preoperative planning of surgical corridor and extent of resection were planned so that maximum possible resection could be achieved without disturbing the WM tracts. DTI indicated the involvement of fiber tracts. A total of 21 (61.7%) patients had a displacement of tracts only and they were not invaded by tumor. A total of 11 (32.3%) patients had an invasion of tracts by the tumor, whereas in 4 (11.7%) patients the tracts were disrupted. Postoperative neurologic examination revealed deterioration of motor power in 4 (11.7%) patients, deterioration of language function in 3 (8.82%) patients, and memory in one patient. Total resection was achieved in 11/18 (61.1%) patients who had displacement of fibers, whereas it was achieved in 5/16 (31.2%) patients when there was infiltration/disruption of tracts. CONCLUSION: DTI provides crucial information regarding the infiltration of the tract and their displaced course due to the tumor. This study indicates that it is a very important tool for the preoperative planning of surgery. The involvement of WM tracts is a strong predictor of the surgical outcome.

Intramedullary Melanocytoma of the Cervicothoracic Cord: Case Report and Review of Literature.

Melanocytoma is rare pigmented tumor of the leptomeninges which arise from the neural crest. Intramedullary location of the tumor is extremely rare, and only a few case reports are available in the literature. We report a case of 35-year-old female with the entity who had a near total removal of the intramedullary tumor with good postoperative outcome. The available literature is reviewed.

Endoscopic partial corpectomy using anterior decompression for cervical myelopathy.

Symptomatic ventral cervical cord compressions have been treated by anterior discectomy with fusion, anterior corpectomy and fusion, discectomy with an artificial disc, and minimally invasive techniques. There are complications associated with the fusion procedure, especially when a long segment fusion is undertaken. Partial removal of the vertebral body without fusion could help in avoiding fusion and its associated complications. We are describing the detailed surgical technique, indications, limitations, and early results of endoscopic partially corpectomy. The detailed endoscopic technique of how to remove minimal portion of the anterior and middle part of the vertebra with the removal of compressing posterior portion is described. To the best of the author's knowledge, this procedure has not been described in the literature till date. This technique has the advantages of preservation of most of the vertebral body and disc. The stability of the spine is maintained without the patient having to undergo the additional procedure of spinal fusion. There is a shorter length of hospital stay. The technique can also be used in special conditions, such as when a migrated disc behind the vertebral body or any bony compression opposite the vertebral body is encountered. It avoids injury to the longus colli muscles and damage to most of the annulus fibrosis. It preserves motion segments at the operated and adjacent segments without any significant decrease in the disc and vertebral height. A relatively short follow-up and the experience in a relatively small number of patients till now, are the limitations to the widespread use of this technique. Although long-term follow-up of this procedure in a large number of patients is not currently available, endoscopic partial corpectomy seems to be an effective and an alternative method for dealing with anteriorly located cervical compression. Long-term follow-up in a large number of patients is required to assess for the safety and effectiveness of the procedure.

Full Endoscopic Vascular Decompression in Trigeminal Neuralgia: Experience of 230 Patients.

BACKGROUND: Although most surgeons are using endoscopy as an adjunct to microscopy in microvascular decompression, a full endoscopic technique is less commonly performed. The present study is aimed to evaluate results of 230 patients of endoscopic vascular decompression. METHODS: A retrospective study was carried out in a tertiary care hospital. Patients with typical neuralgia, with or without preoperatively detected vascular compression, were advised to undergo vascular decompression. RESULTS: Maxillary and mandibular division were involved in 116 and 93 patients, respectively. Superior cerebellar (n = 174) artery was most common vascular conflict followed by anterior inferior cerebellar artery (n = 96). Tortuous basilar artery and small veins were possible causes of neuralgia in 1 and 2 patients, respectively. Single- and double-vessel conflict were observed in 173 and 50 patients, respectively. The compressing vessel was placed anterior to the trigeminal nerve in 39 patients. An arterial loop was in contact with the nerve, producing grooving, and displacing the nerve in 215, 35, and 21 patients, respectively. Complete, satisfactory, and no relief of pain were observed in 204 (88.7%), 11 (5.8%), and 15 (6.5%) patients, respectively. Recurrence was observed in 25 patients at an average follow-up of 60 months. Temporary complications included trigeminal dysesthesia, vertigo, facial paresis, CSF leak, and reduced hearing in 9, 8, 8, 7, and 3 patients, respectively. CONCLUSION: Endoscopic vascular decompression is a safe and efficient alternative technique to endoscopic assisted microvascular decompression provided surgeon is experienced in endoscopic surgery. It is helpful in identification of all offending vessels including the double vessel, and anterior compression without brain and nerve retraction.

Indigenous Inexpensive Practice Models for Skill Development in Neuroendoscopy.

INTRODUCTION: Neurosurgery is a branch having a tough learning curve. Residents generally get very less hands-on exposure for advanced procedures like neuroendoscopy. With the limited number of cadavers available and ethical issues associated with animal models, practice models, and simulators are becoming the able alternative. Most of these simulators are very costly. We tried to build indigenous inexpensive practice models that can help in developing most of the skills of neuroendoscopy. MATERIALS AND METHODS: Models were built for learning hand-eye coordination, dexterity, instrument manipulation, cutting, fine dissection, keyhole concept, drilling, and simulation of laminectomy and ligamentum flavum resection. These were shown in the neuroendoscopic fellowship program conducted in authors' institute, and trainees' responses were recorded. RESULTS: Both novice and experienced neuroendoscopic surgeons validated the models. There was no significant difference between their responses (P = 0.791). CONCLUSION: Indigenous innovative models can be used to learn and teach neuroendoscopic skills. The presented models were reliable, valid, eco-friendly, highly cost-effective, portable, easily made and can be kept in one's chamber for practicing.

Endoscopic technique for single-stage anterior decompression and anterior fusion by transcervical approach in atlantoaxial dislocation.

Although posterior approaches are being used frequently in most atlantoaxial dislocations (AAD), anterior decompression is also required in some patients in whom the C1-2 dislocation is not properly reduced by the posterior approach. Transnasal and transoral approaches need an additional posterior approach to perform atlantoaxial fusion. They also have an added risk of infection. The endoscopic transcervical approach can be used for single-stage cervical decompression and stabilization that includes an odontoidectomy and anterior fusion. It can be used both in reducible and irreducible AAD. Patients with a high basilar invasion, traumatic or other lesions involving the C1 or C2 facet joint, reducible AAD with Chiari malformation, and patients with a large mandible or a mandible angle lying below the C3 level even after the maximum neck extension, should not be subjected to this procedure. Preoperative X-ray, computed tomography (CT) scan with angiogram, and magnetic resonance imaging of the craniovertebral region should be done to assess the dislocation. The early results of an endoscopic transcervical approach were found to be safe and effective for decompression and fusion in our experience. There was no permanent complication. The procedure avoids a two-stage surgery; thus, odontoidectomy, if needed, can be performed in addition to the C1-2 fusion in a single stage.